Single Molecule Analysis of MAGUK Structure and Ligand Binding

MAGUK 结构和配体结合的单分子分析

基本信息

批准号：
10360516
负责人：
Mark E Bowen
金额：
$ 44.49万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-01-10 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10360516
关键词：
Active Sites Address Affect Affinity Allosteric Regulation Award Binding Brain Clinical Cytoplasmic Tail Data Deposition Development Disease Epilepsy Event Family Filament Future Genetic Glutamate Receptor Glutamates In Vitro Individual Kinetics Knowledge Learning Length Ligand Binding Ligands Link Lipids Liquid substance Location Mediating Membrane Memory Mental disorders Methods Modification Molecular Molecular Conformation Mutation Nerve Degeneration Neurosciences Neurotransmitter Receptor Oils Organelles Outcome Peptides Phase Phase Transition Phosphorylation Phosphotransferases Physiological Play Post-Translational Protein Processing Post-Translational Regulation Protein Isoforms Protein Kinase Proteins Proteomics Publishing Reaction Receptor Signaling Role Scaffolding Protein Schizophrenia Sequence Homology Signal Transduction Signaling Protein Site Specificity Stroke Structural Models Structure Synapses Synaptic plasticity System Testing Translations Water Work autism spectrum disorder druggable target glutamatergic signaling in vivo interest membrane activity membrane-associated guanylate kinase nervous system disorder neuropsychiatric disorder neurotransmission novel palmitoylation particle post stroke receptor receptor binding reconstitution recruit scaffold single molecule

项目摘要

This proposal focuses on post-synaptic glutamate signaling, which mediates excitatory neurotransmission. Glutamate receptor signaling is organized by the membrane-associated guanylate kinase (MAGuK) family of scaffold proteins. Scaffolds determine the outcome of signal transduction by controlling the location of neurotransmitter receptors and connecting them to downstream effectors. In particular, two homologous MAGuKs (PSD-95 and PSD-93) play opposing roles in synaptic plasticity yet the basis for their differential activity remains a mystery. A key factor in synaptic plasticity is post-translation modification (PTM) of the MAGuK proteins, which have been missing in most studies that have been published to date. This proposal investigates the posttranslational regulation of PSD-95 and PSD-93. Aim 1 will test the hypothesis that phosphorylation alters the structure and activity of MAGuKs. We propose that a difference in the number and location of PTM sites elicits activity differences. Aim 2 will test the hypothesis that palmitoylation, a lipid modification essential for synaptic targeting, alters MAGuK activity and coverts them into the filament observed at synapses. Studies to date focused on soluble proteins but palmitoylation is indispensable for activity. Aim 3 will test the hypothesis that phase transitions of MAGuKs regulate availability for receptor binding. SynGAP, an essential synaptic protein, induces to liquid phase separation of PSD-95. This can generate a dynamic “membrane-less organelle”. We will investigate the functional effects of phase separation on receptor binding and structure. We possess the only working reconstitution of scaffold interactions in the post-synapse and provided the only kinetic description of MAGuK scaffolding activity. We will use our unique expertise to address how PTM changes the activity of MAGuKs. We have pioneered novel methods for structural refinement of proteins containing intrinsic disorder and generated the only structural model of a full-length scaffold protein to date. We will use these approaches to provide a mechanistic understanding of how MAGuK structure affects scaffolding. Our published work suggests that proteins residing together on PSD-95 have “higher-order” interactions driven by the enforced proximity. Describing the molecular events in excitatory signaling is a fundamental challenge in neuroscience with direct relevance to brain development, memory and learning, and many neurological and neuropsychiatric disorders.

该提案的重点是突触后谷氨酸信号传导，介导兴奋性神经传递。谷氨酸接收器信号传导由膜相关组织鸟苷酸激酶（Maguk）脚手架蛋白家族。脚手架决定了通过控制神经递质接收器的位置并连接来转导信号转导它们对下游效应。特别是两个同源的Maguk（PSD-95和PSD-93）在突触可塑性中扮演相反的角色，但其差异活动的基础仍然是神秘。突触可塑性的关键因素是Maguk的翻译后修饰（PTM）蛋白质，在迄今已发表的大多数研究中都缺少蛋白质。该提案调查了PSD-95和PSD-93的翻译后调节。目的 1将检验以下假设：磷酸化会改变玛格克斯的结构和活性。我们建议PTM站点的数量和位置差异引起活动差异。 AIM 2将检验棕榈酰化的假设，棕榈酰化是突触的必不可少的脂质修饰靶向，改变Maguk活动，并将其覆盖到突触下观察到的细丝中。迄今为止的研究集中在固体蛋白上，但棕榈酰化对于活性是必不可少的。目的 3将检验以下假设，即Maguks的相转换调节接收器的可用性结合。 Syngap是一种必不可少的合成蛋白，诱导PSD-95的液相分离。这可以产生动态的“无膜细胞器”。我们将研究功能相分离对受体结合和结构的影响。我们拥有唯一的脚手架相互作用的重组并提供了Maguk脚手架活动的唯一动力学描述。我们将使用我们的独特专业知识以解决PTM如何改变Maguks的活动。我们有开创性的小说含有内在疾病的蛋白质结构细化的方法，并产生了唯一的迄今为止，全长支架蛋白的结构模型。我们将使用这些方法提供对Maguk结构如何影响脚手架的机械理解。我们出版了工作表明，驻留在PSD-95上的蛋白质具有“高阶”相互作用通过强制接近。描述兴奋信号传导中的分子事件是神经科学中的基本挑战与大脑发育，记忆和直接相关学习，以及许多神经和神经精神疾病。

项目成果

期刊论文数量（25）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Domain orientation in the N-Terminal PDZ tandem from PSD-95 is maintained in the full-length protein.

DOI：
10.1016/j.str.2011.02.017
发表时间：
2011-06-08
期刊：
STRUCTURE
影响因子：
5.7
作者：
McCann, James J.;Zheng, Liqiang;Chiantia, Salvatore;Bowen, Mark E.
通讯作者：
Bowen, Mark E.

Integrative structural dynamics probing of the conformational heterogeneity in synaptosomal-associated protein 25

突触体相关蛋白 25 构象异质性的综合结构动力学探测

DOI：
10.1016/j.xcrp.2021.100616
发表时间：
2021
期刊：
Cell Reports Physical Science
影响因子：
8.9
作者：
Saikia, Nabanita;Yanez-Orozco, Inna S.;Qiu, Ruoyi;Hao, Pengyu;Milikisiyants, Sergey;Ou, Erkang;Hamilton, George L.;Weninger, Keith R.;Smirnova, Tatyana I.;Sanabria, Hugo
通讯作者：
Sanabria, Hugo

Structure and conformational dynamics of Clostridioides difficile toxin A.

DOI：
10.26508/lsa.202201383
发表时间：
2022-06
期刊：
Life science alliance
影响因子：
4.4
作者：
Chen B;Basak S;Chen P;Zhang C;Perry K;Tian S;Yu C;Dong M;Huang L;Bowen ME;Jin R
通讯作者：
Jin R

Beyond the random coil: stochastic conformational switching in intrinsically disordered proteins.

DOI：
10.1016/j.str.2011.01.011
发表时间：
2011-04-13
期刊：
STRUCTURE
影响因子：
5.7
作者：
Choi, Ucheor B.;McCann, James J.;Weninger, Keith R.;Bowen, Mark E.
通讯作者：
Bowen, Mark E.

Site-Specific Phosphorylation of PSD-95 PDZ Domains Reveals Fine-Tuned Regulation of Protein-Protein Interactions.

PSD-95 PDZ 结构域的位点特异性磷酸化揭示了蛋白质-蛋白质相互作用的微调调节。

DOI：
10.1021/acschembio.7b00361
发表时间：
2017
期刊：
ACS chemical biology
影响因子：
4
作者：
Pedersen,SørenW;Albertsen,Louise;Moran,GriffinE;Levesque,Brié;Pedersen,StineB;Bartels,Lina;Wapenaar,Hannah;Ye,Fei;Zhang,Mingjie;Bowen,MarkE;Strømgaard,Kristian
通讯作者：
Strømgaard,Kristian